Catalog # SK6010

Sphingosine Kinase 1 Phospho-Regulation Antibody Kit

Antibody Sampler Kit

DATASHEET  




$395

Sphingolipids are metabolized into bioactive products that include ceramide, sphingosine, and sphingosine-1-phosphate. SK catalyzes the phosphorylation of the lipid sphingosine, creating the bioactive lipid sphingosine-1-phosphate (S1P). S1P subsequently signals through cell surface G protein-coupled receptors, as well as intracellularly, to modulate cell proliferation, survival, motility, and differentiation. Two isoforms of SK have been identified, SK1 and SK2. The mRNA for both of these isoforms is widely expressed with SK1 expression highest in brain, heart, kidney, thymus, spleen, and lung, while SK2 is highest in kidney and liver. SK can be activated through growth factor, G protein-coupled, and immunoglobulin receptor signaling. SK1 has been shown to mediate cell growth, prevention of apoptosis, and cellular transformation, and is upregulated in a variety of human tumors. Regulation of SK1 may occur through ERK mediated phosphorylation of Ser-225. This phosphorylation leads to increased activity and translocation to the plasma membrane.

References
Melendez, A.J. et al. (2000) Gene 251(1) :19.
Pitson, S.M. et al. (2003) EMBOJ. 22(20) :5491.
Pitson, S.M. et al. (2005) J Exp. Med. 201(1) :49.

Western blot image of 47 kDa recombinant his-tagged human SK1 protein that was phosphorylated with ERK2. The blots were probed with anti-SK1 (Central region) (SP1621; lanes 1-3) or anti-SK1 (Ser-225) (SP1641; lanes 4-6). Both antibodies where used in the presence of no peptide (lanes 1 & 4), phospho-SK1 (Ser-225) peptide (SX1645; lanes 2 & 5), or unphosphorylated SK1 (Ser-225) peptide (SX1625; lanes 3 & 6).

Western blot image of 47 kDa recombinant his-tagged human SK1 protein that was phosphorylated with ERK2. The blots were probed with anti-SK1 (Central region) (SP1621) at 1:500 (lane 1), 1:1000 (lane 2), and 1:2000 (lane 3), or or anti-SK1 (Ser-225) (SP1641) at 1:250 (lane 4), 1:500 (lane 5), and 1:1000 (lane 6).



Rabbit polyclonal antibodies are supplied in phosphate-buffered saline (PBS), 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. The secondary reagents are supplied in the same buffer without azide, and the peptides are supplied in PBS and 0.05% sodium azide. Store all at –20°C. Stable for 1 year.

The products are are safely shipped at ambient temperature for both domestic and international shipments. Each product is guaranteed to match the specifications as indicated on the corresponding technical data sheet. Please store at -20C upon arrival for long term storage.



*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.

Product References:

SP1641 Xiu, L. et al. (2015) Am J Pathol. 185(2):387 WB: hMSC cells
SP1641 Durham, J. et al. (2015) Invest Ophthal Vis Sci. 56(6):3441 WB: human retinal cells and pericytes
SP1641 Sun, K. et al. (2015)Blood 125(10): 1643. WB: mouse erythrocyte
SP1641 Wilson, P.C. et al. (2015) Mol Endocrinol. 29(6):896 WB: rat aortic vascular smooth muscle cells
SP1641 Kim, E.Y. et al. (2014) FASEB J. 28(10): 4347 WB: HEK293-CD40 cells
SP1641 Yasuo, M. et al. (2013) PLoS One 8(1):e53927. WB: rat lung
RS3251 Kawasaki, H. et al. (2013) World J Gastroenter. 19(17):2629. WB, ICC: mouse intestinal myofibroblasts
SP1641 Dai, L. et al. (2013) Molec Endocrinology 28(2):197. WB: human MCF7 breast cancer
SP1641 Park, K. et al. (2013) Mol Cell Biol. 33(4)752. WB: human keratinocytes
SP1621 Chakraborty, G. et al. (2012) J Neurochem. 121(5):806. WB: rat forebrain
SP1621 Yang, L. et al. (2012) Am J Pathol. 181(1):85. WB, ICC: mouse bone marrow stem cells
RS3251 Estrada-Bernal, A. et al. (2011) J Neurooncol. 102:353. Western Blot
SP1641 Alvarez, S.E. et al. (2010) Nature 465:1084. WB: A7 melanoma cells
SP1641 Miller et al. (2008) Mol Cell Biol 28(12):4142. WB: human esophageal carcinoma
SP1641 Paugh, B.S. et al. (2008) FASEB J. 22(2):455. WB: human glioblastoma A172
SP1641 Shida, D. et al. (2008) Cancer Res. 68(16):6569. WB: human MNK1 cells + LPA
SP1621 Tao, R. et al. (2007) Cardiovas. Res. 74:56. WB: mouse heart, wildtype vs null

This kit contains:

CATALOG#DESCRIPTIONSIZEAPPLICATIONSSPECIES REACTIVITYMW (kDa)
Sphingosine Kinase 1 (Central region) Rabbit pAb50 μlWB, EHu47
Sphingosine Kinase 1 (Ser-225), phospho-specific Rabbit pAb50 μlWB, EHu47
phospho-Sphingosine Kinase 1 (Ser-225) Peptide50 μgAB, E
unphosphorylated Sphingosine Kinase 1 (Ser-225) Peptide 50 μgAB, E
Anti-Rabbit Ig Light-Chain Specific:HRP Mouse mAb100 μlWB, E, ICC, IHCRb
KIT SUMMARY

The sphingosine kinase 1 (SK1) phospho-regulation antibody sampler kit can be used to detect the level of phosphorylation of SK1 (Tyr-225) relative to the total expression level of SK1. The kit also includes peptides for antibody blocking experiments and a secondary reagent for antibody detection.